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The  Dissertation Service  accepts the approved version of dissertations and post-doctoral theses (habilitations) from the TU Berlin and checks compliance with the formal and technical specifications. The team publishes the works, enters them into the appropriate bibliographic databases and issues the official acknowledgments of receipt.

For all other further information on doctoral degrees please contact your respective  Faculty-Service-Center  or the  Center for Junior Scholars (CJS, formerly: TU-DOC Office for Doctoral and Postdoctoral Services) at TU Berlin.

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According to § 9 (2) of the Regulations for Doctorates at TU Berlin , the scientific community must be given appropriate access to a dissertation after completing the scientific defense. 

"Access" means that the version approved by the Doctoral Committee must be delivered to the University Library free of charge in one of the following forms to ensure proper dissemination:

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If the habilitation regulations of your Faculty require the post-doctoral thesis to be submitted to the University Library, please follow the checklist below: Checklist for publishing a habilitation thesis

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2024 Theses Doctoral

Non-viral delivery of nucleic acid gene editing components to the liver and brain

Cai, Shuting Sarah

In the growing landscape of innovative non-viral delivery vehicles, polymeric and lipid nanoparticles remain at the forefront for their versatility in encapsulating a variety of therapeutic payloads. This thesis investigates their potential for facilitating the transport of nucleic acid components into cells, with a focus on targeted delivery to the liver and brain. To achieve this, we address key considerations including the composition of the delivery vector, the nature of the therapeutic cargo, and the chosen delivery route. The challenge of targeted delivery to specific organs or cell types, i.e. hepatocytes or neurons, is addressed through rational design and development of libraries of nanoparticulate systems tailored for nucleic acid therapeutics. Although liver gene editing using non-viral systems has been extensively studied, oral delivery for liver targeting remains challenging due to the mucosal barrier. To that end, we explore intraduodenal delivery as a strategy to bypass the mucosal barrier and target the liver. Furthermore, insights from collaborative research with the Mao lab at Johns Hopkins University reveal that tuning the composition of lipid nanoparticles (LNPs) can influence their preferential targeting of specific cell types. Leveraging this, we employed an in vitro library screening and machine learning approach to identify populations of LNPs capable of preferentially transfecting hepatocytes. The efficacy of these LNPs in liver gene editing is then evaluated through “cluster-mode” screening in vivo, and therapeutic efficacy was demonstrated using a proof-of-concept in vivo model for PCSK9 and ANGPTL3 knockdown, resulting in 27% serum cholesterol knockdown. In addition to liver-targeted gene delivery, this thesis also investigates the potential of polymeric and lipid nanoparticles for delivering nucleic acid therapeutics to the brain. However, overcoming the blood-brain barrier (BBB) is crucial for systemic delivery to the brain. To circumvent the BBB, we explored two methods: intracranial injection and theranostic ultrasound (THUS)-mediated temporary opening of the BBB. While intracranial injection achieves localized gene editing, THUS offers a non-invasive approach for transient and widespread BBB opening. Utilizing the previously validated in vitro screening and machine learning approaches for chitosan-grafted bPEI (CS-PEI) and lipid nanoparticle (LNP) carriers with tunable compositions, we assessed their efficacy in systemic gene delivery to the brain, and specifically their capability in preferentially transfecting neuronal cells over hepatocytes. Subsequently, we validated their efficiency via intracranial administration using the Ai14 reporter mouse model and observed up to 20% gene editing of the targeted cross-sectional area of the brain hemisphere using the top-performing cluster. Through comprehensive investigations into both brain and liver gene delivery, this thesis aims to contribute to the advancement of non-viral nanoparticle-based gene therapy strategies for treating a range of cholestatic liver diseases and hereditary neurodegenerative diseases.

• Biomedical engineering
• Nucleic acids
• Gene editing
• Nanoparticles
• Machine learning
• Liver--Diseases--Treatment
• Johns Hopkins University

This item is currently under embargo. It will be available starting 2029-04-25.

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